Utilització per primera vegada de metabolits per valoara l'impacte del fum de tercera mà en l'exposició al tabac en no fumadors

1. THIRDHAND SMOKE: a new pathway of
passive smoking exposure
Noelia Ramírez, PhD
Metabolomics Platform-IISPV-URV
H2020-MSCA-IF-2014 - 660034 - THSExposure
Tobacco Control Seminars
14th June 2016

2. Tobacco Control Seminars. UCT-ICO, 14th June 2016 2
Outline
• What is Thirdhand Smoke (THS)?
• THS: Advances on risk assessment & health impact
• Why metabolomics in THS research?
• THS & metabolomics: first studies
• Conclusions

3. Tobacco Control Seminars. UCT-ICO, 14th June 2016 3
What is Thirdhand Smoke (THS)?
Second hand smoke (SHS)

4. Tobacco Control Seminars. UCT-ICO, 14th June 2016 4
Thirdhand
Smoke
(THS)
SHS/ETS
Nicotine
TSNAs
N
N
O N
O
N
O
Volatile N-nitrosamines
Amines
Nitro
aromatics
What is Thirdhand Smoke (THS)?

5. Tobacco Control Seminars. UCT-ICO, 14th June 2016 5
IARC classification
Group 1: Carcinogenic to
humans
Group 3: Not classifiable as to
its carcinogenicity to
humans
Tobacco Specific Nitrosamines (TSNAs)
Nitrosation HONO (NO+)
NNK
Reduction
NNAL
Demethylation Nitrosation
NNN
NAB
NAT
Nicotine
NNA
Oxidation
iso-NNAL
Reduction
iso-NNAC
Exclusive in THS
What is Thirdhand Smoke (THS)?

6. Tobacco Control Seminars. UCT-ICO, 14th June 2016 6
What is Thirdhand Smoke (THS)?

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Pediatrics 123 (1) 2009, e74-e79
National survey (%, N=1478)
Believe that SHS harms children
Agree 93.2
Disagree 3.3
Don't know 3.4
Believe that THS harms children
Agree 61.0
Disagree 16.7
Don't know 22.3
Smoking parents beliefs (%, N=1770)
Believe that THS harms children
Agree 91
Disagree 9
Mothers > Fathers
Smokers (<10 cigarettes/day)
What is Thirdhand Smoke (THS)?

8. Tobacco Control Seminars. UCT-ICO, 14th June 2016 8
POSED RULE IS PENDING PUBLICATION IN THE FEDERAL REGISTER. PUBLICATION
THE FEDERAL REGISTER WILL COMMENCE THE PUBLIC COMMENT PROCESS
DEPARTMENT OF HOUSING AND URBAN DEVELOPMENT
24 CFR Parts 965 and 966
[Docket No. FR 5597-P-02]
RIN 2577-AC97
Instituting Smoke-Free Public Housing
NCY: Office of the Assistant Secretary for Public and Indian Housing, HUD.
ON: Proposed rule.
MARY: This proposed rule would require each public housing agency (PHA)
Proposed rule. USA Federal Government.
22nd October 2015
Env. Health Perspect. 119 (2) 2011, A70-A74 Env. Health Perspect. 119 (2) 2011, 1218-1226
Consortium on Thirdhand Smoke
What is Thirdhand Smoke (THS)?

9. Tobacco Control Seminars. UCT-ICO, 14th June 2016 9
Thresholds:
WHO: 10-5
USEPA: 10-6
Lifetime cancer risk, by group age
Non-smokers’ house dust Smokers’ house dust
THS: Advances on risk assessment & health impact

10. Tobacco Control Seminars. UCT-ICO, 14th June 2016 10
Environ. Sci. Technol. 2014, 48,13093
DALYs for non-smokers living with smokers
(4 possible scenario)
Annual DALYs for non-smokers living with
smokers
DALYs
Disability-Adjusted Life
Years
THS: Advances on risk assessment & health impact

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THS: Advances on risk assessment & health impact

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Cancer Epidemiol Biomarkers Prev 20 (6) 2011, 1213-1221
Cotinine in non-
smokers’ urine
Cotinine & NNAL in urine
from children exposed to
SHS & THS
THS: Advances on risk assessment & health impact
Population-based studies

13. Tobacco Control Seminars. UCT-ICO, 14th June 2016 13
Mutagenesis, 28 (2014) 381
THS: Advances on risk assessment & health impact
• DNA damage in THS exposed cells
• Increased oxidative stress
• THS is genotoxic in human cell
lines
Consortium on Thirdhand Smoke

14. Tobacco Control Seminars. UCT-ICO, 14th June 2016 14
Plos One, 9 (2014) e86391
THS-exposed mice:
• Dyslipidaemia
• Hyperglycaemia
• Hyperactivity and risk of
neurological disorders
• Poor wound healing
Lungs
• Inflammation and excess
of pro-inflammatory
cytokines/chemokines
• Elevated levels of collagen
fibres.
THS: Advances on risk assessment & health impact
Consortium on Thirdhand Smoke

15. Tobacco Control Seminars. UCT-ICO, 14th June 2016 15
THS-exposed mice:
• Visceral fat accumulation
• Increased fasting blood
glucose
• Fasting serum insulin
• Reduced protein levels of the
skeletal muscle (IR, PI3k &
AKT)
• Oxidative stress in the
skeletal muscle
THS: Advances on risk assessment & health impact
Plos One, 11 (2016) e0149510
Effects increased by
high-fat diet
Effects decreased by
antioxidants

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• Incomplete characterisation of THS composition and mechanisms
of formation
• Few remediation studies
• Health risk assessment based on the available toxicological data
• Lack of population-based studies
THS: current uncertainties and main challenges
THS: Advances on risk assessment & health impact
METABOLOMICS

17. Tobacco Control Seminars. UCT-ICO, 14th June 2016 17
Why metabolomics in THS research?
"Genomics and proteomics tell you what might happen, but metabolomics tells you what
actually did happen." , Bill Lasley, University of California (UC), Davis.
Genomics
Transcriptomics
Proteomics
Metabolomics

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Targeted Metabolomics
Focused on specific metabolites:
• typically hypothesis-driven
• effect of drug/disease on specific pathway
• often performed with QqQ in MRM mode
Why metabolomics in THS research?

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Why metabolomics in THS research?
Untargeted Metabolomics
Changes to any metabolite(s) within
the entire metabolome:
• Typically not hypothesis-driven
• Measuring, ideally, the largest set
of metabolites at the raw signal
level and in an unbiased manner
• Changes to any metabolite(s) within
the entire metabolome
• Investigations into unknown
mechanisms
Main analytical challenges:
• Complexity of biological matrices
• Hundreds to thousand compounds
with high chemical variety at a wide
range of different concentrations.

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Why metabolomics in THS research?
Experimental
design
Untargeted metabolomics workflow

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Sci Rep. 2016; 6:23849.
Consortium on Thirdhand Smoke
THS & metabolomics: first studies

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C57 mice THS-exposed
for 6 months Biofluids: serum & urine
Untargeted metabolomics by GC-QToF &HPLC-QToF
Plos ONE, 9 (2014)1:e86391.
Metabolites extraction
Derivatization
GC-QToF analysis
Prof. Martins-Green
Cell Biology & Neuroscience
THS & metabolomics: first studies

23. Tobacco Control Seminars. UCT-ICO, 14th June 2016 23
Urine
THS exposed
Control
Sugars
THS & metabolomics: first studies
Serum
THS exposed
Control
Sugars
Lipids

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Alanine
Aspartic acid
Glucose
Glucuronic acid
Glutamate
Glycerol
Isoleucine
Leucine
Methionine
Ornithine
Phenyl alanine
Proline
Pseudo uridine
Serine
Tryptophan
Threonine
Valine
Phospholipids…
Fatty acids…
Differential metabolites that match with
tobacco & metabolomics studies
Putative identifications
THS & metabolomics: first studies

25. Tobacco Control Seminars. UCT-ICO, 14th June 2016 25
THS & metabolomics: first studies
Urine
C57 mice THS-exposed for 6 months
Four groups: Control
THS
THS+WD
THS+Antiox.
967 differentially expressed peaks (p<0.05; FC>2; HMDB hits )

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THS & metabolomics: first studies
Ctrl. vs THS. Enrichment results
Affected metabolic pathways
• Tryptophan Metabolism
• Arginine and Proline Metabolism
• Arachidonic Acid Metabolism
• Tyrosine Metabolism
• Methionine Metabolism
• Phenylalanine metabolism
• D-Glutamine and D-glutamate metabolism
• Histidine metabolism
• Phenylalanine, tyrosine and tryptophan biosynthesis
• Alanine, aspartate and glutamate metabolism
• Vitamin B6 metabolism
• Cyanoamino acid metabolism
• Steroid hormone biosynthesis
• …
Putative identifications
Preliminary results

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KMO
IDO1
Glucuronide
KAT
THS & metabolomics: first studies
Putative identifications
Preliminary results

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Mass Spectrometry Imaging by LDI-TOF
THS & metabolomics: first studies

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Mass Spectrometry Imaging by LDI-TOF
THS & metabolomics: first studies

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Conclusiones
• Thirdhand smoke (THS) exposure is a health risk underestimated by
the general population and tobacco control policies.
• Several studies on risk assessment, THS exposure of human cell
lines and animal models presented evidences of the health risks
associated to this exposition.
• Future research should focus on finding more evidences of health
effects, especially on population-based studies.
• The use of metabolomics would allow the identification of the
metabolic changes associates to THS exposure.
• Preliminary results in animal models are really promising, suggesting
that metabolomics would imply a giant step on the evaluation of
tobacco consumption and THS/SHS exposure, particularly.

31. Tobacco Control Seminars. UCT-ICO, 14th June 2016 31
[@] noelia.ramirez@urv.cat
[www] www.metabolomicsplatform.com
Prof. Martins-Green
Cell Biology & Neuroscience
California THS Consortium
Agradecimientos
Metabolomics Platform
Contacto:
This project has received funding from the European Union’s Horizon 2020 research and
innovation programme under the Marie Sklodowska-Curie grant agreement No. 660034.

32. TABAQUISMO DE TERCERA MANO
Thirdhand tobacco smoke (THS)
Dra. Noelia Ramírez
Metabolomics Platform-IISPV-URV
H2020-MSCA-IF-2014 - 660034 - THSExposure
XIX REUNIÓN DE INVIERNO SEPAR.
ÁREA DE TABAQUISMO
Alcalá de Henares, 20 noviembre
2015